TARGETING AND FUNCTION OF CENTROMERE PROTEIN A

染色体分离在有丝分裂和减数分裂是必不可少的所有 细胞繁殖 这一进程中的中断具有深远的影响， 对人类健康的后果。 着丝粒指定装配 染色质纤维上一组独特的功能元件， 提供运动性、姐妹染色单体凝聚力和机械感觉 将这些功能与细胞周期检查点联系起来的机制。 CENP-A是一种着丝粒特异性的核心同源物， 核小体蛋白组蛋白H3。 CENP-A的特异性组装 需要蛋白质-蛋白质 核小体内的相互作用，分化的DNA接触位点 在核小体的表面和一个独特的调节途径， 从而提出了选择性识别着丝粒 CENP-A组蛋白复合物的DNA。 CENP-A核小体将 用亲和层析分离的着丝粒DNA序列重建 对于CENP-A。 DNA-蛋白质相互作用的分析将揭示 CENP-A是否表现出特异性识别着丝粒DNA。 将检查CENP-A组装到染色质中以进行测试 CENP-A染色质装配通过一个 新的非S期途径。 CENP-A相关蛋白将被 分离并表征以探索其功能相互作用。 突变方法将结合联合收割机长期转染方法 通过新颖的着丝粒运动性测定方法、机械 活细胞中的特性和空间组织。

Chromosome segregation at mitosis and meiosis is essential for all cellular reproduction. Disruptions in this process have profound consequences for human health. Centromeres specify the assembly of a unique set of functional elements on the chromatin fiber that provide motility, sister chromatid cohesion and mechanosensory mechanisms that link these functions to cell cycle checkpoints. CENP-A is a centromere specific homologue of the core nucleosomal protein histone H3. The specific assembly of CENP-A at human centromeres requires a combination of protein-protein interactions within the nucleosome, differentiated DNA contact sites at the surface of the nucleosome and a unique regulatory pathway, leading to a hypothesis for selective recognition of centromeric DNA by CENP-A histone complexes. CENP-A nucleosomes will reconstituted with centromeric DNA sequences isolated by affinity for CENP-A. Analysis of DNA-protein interactions will reveal whether CENP-A exhibits specific recognition of centromeric DNA. The assembly of CENP-A into chromatin will be examined to test the hypothesis that CENP-A chromatin assembly occurs through a novel, non-S phase pathway. CENP-A associated proteins will be isolated and characterized to explore its functional interactions. Mutational approaches will combine long term transfection methods with novel assay methods for centromere motility, mechanical properties and spatial organization in living cells.